Gregory S P, Dillon N O, Butterworth P H
Nucleic Acids Res. 1982 Dec 11;10(23):7581-92. doi: 10.1093/nar/10.23.7581.
The mRNA start site of a cloned rainbow trout protamine gene (TPG-3) has been localised using S1-nuclease mapping and primer extension of in vivo synthesised trout testis poly A+-RNA. The presumptive cap site occurs within an AT-rich region, only 14 nucleotides from the start of the protein-coding sequence. Transcription of this protamine gene in vitro, using the Hela whole-cell extract system, generates products initiated at the same nucleotide as that used in vivo. In vitro transcription is abolished by deletion of sequences between -20 and -48, within which is a canonical TATA-box having an llbp homology with the strong chick conalbumin and Adenovirus-2 major late promoters (CTATAAAAGGG).
利用S1核酸酶图谱分析和体内合成的虹鳟鱼睾丸聚腺苷酸加尾RNA的引物延伸法，已确定了克隆的虹鳟鱼鱼精蛋白基因（TPG - 3）的mRNA起始位点。推定的帽位点位于富含AT的区域内，距离蛋白质编码序列起始处仅14个核苷酸。使用Hela全细胞提取物系统在体外转录该鱼精蛋白基因，产生的产物起始于与体内相同的核苷酸。通过缺失-20至-48之间的序列可消除体外转录，该区域内有一个典型的TATA框，与强的鸡伴清蛋白和腺病毒2型主要晚期启动子（CTATAAAAGGG）具有11bp的同源性。
